Sladek T L, Jacobberger J W
Department of Genetics, Case Western Reserve University, School of Medicine, Cleveland, Ohio 44106.
Cytometry. 1993;14(1):23-31. doi: 10.1002/cyto.990140106.
Few quantitative studies addressing immunofluorescence histogram analysis have been published. One study by Overton (Cytometry 9:619-626, 1988) has shown threshold and histogram subtraction methods to be accurate for analysis of well-separated immunofluorescence distributions of positive and negative cells. An evaluation of methods to analyze immunofluorescence histograms when positive and negative immunofluorescence distributions overlap has not, to our knowledge, been reported. In this paper, data obtained from flow cytometry of immunofluorescently stained cells infected with recombinant retroviruses that produce a range of simian virus 40 large T antigen levels were analyzed by threshold, histogram subtraction, and distribution modeling methods. This analysis showed that as the separation between the immunofluorescence distributions of positive and negative cell populations decrease the best methods for histogram analysis are modeling followed, in order, by histogram subtraction, and threshold analysis.
很少有关于免疫荧光直方图分析的定量研究发表。奥弗顿的一项研究（《细胞计数》9:619 - 626，1988年）表明，阈值法和直方图减法对于分析阳性和阴性细胞分离良好的免疫荧光分布是准确的。据我们所知，尚未有关于阳性和阴性免疫荧光分布重叠时免疫荧光直方图分析方法的评估报道。在本文中，通过阈值法、直方图减法和分布建模方法，对感染产生一系列猿猴病毒40大T抗原水平的重组逆转录病毒的免疫荧光染色细胞进行流式细胞术获得的数据进行了分析。该分析表明，随着阳性和阴性细胞群体免疫荧光分布之间的间距减小，直方图分析的最佳方法依次为建模法、直方图减法和阈值分析法。
